中药DNA条形码鉴定中的DNA提取方法研究

摘要

目的:建立一种直接从药材中提取DNA的方法以满足中药材DNA条形码研究的需要.方法:以麦冬药材为实验对象,对比5种常用植物DNA提取方法,筛选出最优方法后,对不同部位入药的10种中药材进行DNA提取,采用琼脂糖凝胶电泳及PCR扩增效果进行检测.结果:改良的CTAB法较适合药材DNA提取,虽然所有样品DNA都出现降解,但基于ITS2序列引物PCR扩增成功率为90％.结论:通过增加药材的预处理、核分离液等步骤,并改良CTAB浓度等措施,CTAB法能够用于上述不同药用部位入药的9种药材DNA提取,可以为中药DNA条形码鉴定中的DNA提取提供参考.%This study was aimed to establish a universal DNA extraction method for future research on authentication of Chinese herbal medicine by DNA barcoding. Ten representative specimens belonging to five different medicinal parts were used to evaluate five different DNA extraction methods. The universal DNA extraction method was further modified and evaluated by agarose gel electrophoresis and PCR amplification. Modified CTAB method was a common extraction method for almost all kinds of herbs. The amplification success rate for ITS2 was 90% though DNA degradation occurred in all specimens. The study showed that modified CTAB method was suitable for nine herbs by improving methods on herbal pretreatment, buffer for DNA separation, and concentration of CTAB. The simple and feasible CTAB approach provided a reference method for DNA extraction of herbal medicines.